Programmable N6-methyladenosine modification of CDCP1 mRNA by RCas9-methyltransferase like 3 conjugates promotes bladder
- PDF / 3,252,000 Bytes
- 6 Pages / 595.276 x 790.866 pts Page_size
- 44 Downloads / 135 Views
LETTER TO THE EDITOR
Open Access
Programmable N6-methyladenosine modification of CDCP1 mRNA by RCas9methyltransferase like 3 conjugates promotes bladder cancer development Xiaoling Ying1,2, Xu Jiang1, Haiqing Zhang1, Bixia Liu1, Yapeng Huang3, Xiaowei Zhu3, Defeng Qi3, Gang Yuan4, Junhang Luo2 and Weidong Ji1*
Abstract Accumulating evidence has revealed significant roles for N6-methyladenosine (m 6 A) modification in the development of various cancers. We previously demonstrated an oncogenic role of m 6 A-modified CUB domain containing protein 1 (CDCP1) in bladder cancer (BC) progression. However, the biological functions and underlying molecular mechanisms of engineered programmable m 6 A modification of CDCP1 mRNA in BC remain obscure. Here, we established a targeted m 6 A RNA methylation system by fusing the catalytic domain of methyltransferase like 3 (METTL3CD) to RCas9 as the RNA-targeting module. The constructed RCas9- METTL3 retained methylation activity and mediated efficient site-specific m 6 A installation in the presence of a cognate single guide RNA and short protospacer adjacent motif-containing ssDNA molecule . Subsequently, targeting m 6 A installation onto the 3′ untranslated region of CDCP1 promoted CDCP1 mRNA translation and facilitated BC development in vitro and in vivo. Our findings demonstrate that the RCas9-METTL3 system mediates efficient sitespecific m 6 A installation on CDCP1 mRNA and promotes BC development. Thus, the RCas9-METTL3 system provides a new tool for studying m 6 A function and a potential strategy for BC epitranscriptome-modulating therapies.
Background RNA epitranscriptomics has gained popularity in recent years [1]. To date, more than 160 different RNA modifications have been identified [2]. Of these, N6methyladenosine (m6A) is the most prevalent RNA modification in eukaryotes [3]. m6A modification is especially relevant to the occurrence and development of tumors. m6A methyltransferases may play oncogenic or suppressive roles in malignant tumors. Methyltransferase like 3 (METTL3) promotes the progression of bladder cancer (BC) by regulating the expression levels of AFF4, * Correspondence: [email protected] 1 Center for Translational Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China Full list of author information is available at the end of the article
IKBKB, RELA, MYC, ITGA6, and CUB domaincontaining protein 1 (CDCP1) or by accelerating premiR221/222 maturation in an m6A-dependent manner [4–7]. However, METTL14 inhibits the self-renewal capacity of BC-initiating cells and bladder tumorigenesis by modulating Notch1 m6A levels [8]. The methyltransferase family proteins recognize their specific sites and modify targeted transcripts differentially, suggesting that the location of the m6A modifications on mRNA transcripts may underlie the observed differences. As knockdown or overexpression of m6A methyltransferases leads to altered m6A content at numerous sites on many transcripts, it is difficult to determine the roles of
Data Loading...
